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目的建立螺旋藻中维生素B1测定的高效液相色谱法。方法试样加0.2mol/L盐酸,充分摇匀,在恒温干燥箱内121℃水解30min后,冷却,用2.0mol/L乙酸钠溶液调pH值至4.5左右,再加木瓜蛋白酶和淀粉酶混合酶液,摇匀,置37℃的培养箱中过夜,纯水定容后过滤,取滤液经碱性铁氰化钾衍生,正丁醇萃取,用symmetry C18(5μm,250mm×4.6mm)柱分离,以甲醇和乙酸钠为流动相,用荧光检测器(Ex:375nm,Em:435nm)检测,外标法定量。结果该方法基线平稳,峰分离完全,检出限为0.2mg/kg,线性范围为0.01~2.0mg/L,加标回收率为86.36%~98.23%,精密度为1.15%~3.14%。结论该方法操作简便、准确可靠,适用于螺旋藻类样本中维生素B1的测定。
Objective To establish a HPLC method for the determination of vitamin B1 in Spirulina. Method The sample was added with 0.2mol / L hydrochloric acid, shake well, in the oven at 121 ℃ hydrolysis 30min, cooled, with 2.0mol / L sodium acetate solution to adjust the pH to 4.5 or so, plus papain and amylase mixed The enzyme solution was shaken and placed in an incubator at 37 ° C overnight. After the pure water was filtered, the filtrate was derivatized with basic potassium ferricyanide and extracted with n-butanol. The filtrate was purified on a Symmetry C18 (5 μm, 250 mm × 4.6 mm) column Separation, methanol and sodium acetate as the mobile phase, with a fluorescence detector (Ex: 375nm, Em: 435nm) detection, external standard method. Results The method was stable and the peak was completely separated. The detection limit was 0.2 mg / kg. The linear range was 0.01-2.0 mg / L. The recoveries were 86.36% -98.23% and the precision was 1.15% -3.14%. Conclusion The method is simple, accurate and reliable and is suitable for the determination of vitamin B1 in spirulina samples.